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Analysis of transcriptional co-regulation and identification of genes of biotechnological interest in Trichoderma reesei

Grant number: 15/08222-8
Support type:Scholarships in Brazil - Doctorate
Effective date (Start): July 01, 2015
Effective date (End): July 04, 2019
Field of knowledge:Biological Sciences - Genetics
Cooperation agreement: Coordination of Improvement of Higher Education Personnel (CAPES)
Principal Investigator:Juliana Velasco de Castro Oliveira
Grantee:Gustavo Pagotto Borin
Home Institution: Centro Nacional de Pesquisa em Energia e Materiais (CNPEM). Ministério da Ciência, Tecnologia, Inovações e Comunicações (Brasil). Campinas , SP, Brazil
Associated scholarship(s):17/18987-7 - Construction of mutant Trichoderma reesei strains and analysis of regulatory elements aiming to identify and characterize new genes with biotechnological potential for lignocellulose deconstruction, BE.EP.DR

Abstract

Plant cell wall is a recalcitrant structure composed of complex polysaccharides which can be broken down into fermentable sugars for the production of second generation (2G) ethanol. The deconstruction of this complex material can be made by a variety of hydrolytic enzymes which are naturally produced by a variety of microorganisms. Among them, the fungus Trichoderma reesei stands out for its ability to produce and secrete these enzymes in large quantities. Although this fungus is widely studied as a model organism in the deconstruction of lignocellulose, only recently transcriptomics and proteomics approaches with T. reesei RUT C30 grown in two plant biomass of sugarcane were carried out. In the master's project recently concluded by the candidate it was evaluated the expression profile and secretion of proteins involved in the deconstruction of plant wall. The results of T. reesei transcriptome allowed the identification of a set of differentially expressed genes in culm and bagasse, which can be related, directly or indirectly, to the deconstruction and fungal growth in such lignocellulosic substrates. Thus, this PhD project aims to continue with this project and study in more detail the results obtained. The objective of this proposal is to analyze the co-regulation of genes differentially expressed in the T. reesei transcriptome analysis inferring a gene network. In addition, it is proposed as objective to construct and validate mutant strains of this fungus to identify genes with unknown function which may be of biotechnological interest as new lignocellulolytic enzymes. Knockout mutants and mutants overexpressing the genes selected will be constructed to evaluate their lignocellulolytic profiles by growth tests in media containing different sources of carbon, measuring enzymatic activity against various substrates and analyzing the amount of sugar released during saccharification, and others. As additional objectives, it is proposed (I) to clone, express, purify and characterize one or more proteins encoded by genes which had the most interesting profiles in the mutant and (II) to analyze the T. reesei metabolomics profile during growth on sugarcane biomass to identify the main metabolic pathways used by the fungus. At the end of this project, it is expected to identify potential genes to increase plant biomass saccharification of sugarcane and the production of 2G ethanol. (AU)

Scientific publications
(References retrieved automatically from Web of Science and SciELO through information on FAPESP grants and their corresponding numbers as mentioned in the publications by the authors)
BONIN, GUSTAVO PAGOTTO; CARAZZOLLE, MARCELO FALSARELLA; CORREA DOS SANTOS, RENATO AUGUSTO; RIANO-PACHON, DIEGO MAURICIO; DE CASTRO OLIVEIRA, JULIANA VELASCO. Gene Co-expression Network Reveals Potential New Genes Related to Sugarcane Bagasse Degradation in Trichoderma reesei RUT-30. FRONTIERS IN BIOENGINEERING AND BIOTECHNOLOGY, v. 6, OCT 22 2018. Web of Science Citations: 1.
BORIN, GUSTAVO PAGOTTO; SANCHEZ, CAMILA CRISTINA; DE SANTANA, ELIANE SILVA; ZANINI, GUILHERME KEPPE; CORREA DOS SANTOS, RENATO AUGUSTO; PONTES, ANGELICA DE OLIVEIRA; DE SOUZA, ALINE TIEPPO; TAVARES SOARES DAL'MAS, ROBERTA MARIA MENEGALDO; RIANO-PACHON, DIEGO MAURICIO; GOLDMAN, GUSTAVO HENRIQUE; DE CASTRO OLIVEIRA, JULIANA VELASCO. Comparative transcriptome analysis reveals different strategies for degradation of steam-exploded sugarcane bagasse by Aspergillus niger and Trichoderma reesei. BMC Genomics, v. 18, JUN 30 2017. Web of Science Citations: 21.
KAUPERT NETO, ANTONIO ADALBERTO; BORIN, GUSTAVO PAGOTTO; GOLDMAN, GUSTAVO HENRIQUE; DE LIMA DAMASIO, ANDRE RICARDO; DE CASTRO OLIVEIRA, JULIANA VELASCO. Insights into the plant polysaccharide degradation potential of the xylanolytic yeast Pseudozyma brasiliensis. FEMS Yeast Research, v. 16, n. 2 MAR 2016. Web of Science Citations: 1.

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